Conventional Isoelectric Focusing

In principle, pH gradients could be obtained by diffusion of non-amphoteric buffers but such 'artificial' gradients would be altered by changes in electric migration and diffusion of the buffer ions. Thus, Svensson in 1961 introduced the concept of 'natural' pH gradients, created and stabilized by the electric current itself. The buffers used in this system required two fundamental properties: first amphoterism, so that they could reach an equilibrium position along the separation column and secondly 'carrier' ability. This last concept is more subtle, but just as fundamental. Any ampholyte cannot simply be used for IEF; only a carrier ampholyte, that is a compound capable of transporting the current (a good conductor) and capable of carrying the pH (a good buffer). With this notion, and with Vesterberg's elegant synthesis of such ampholytes in 1969, present-day conventional IEF was born.

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